Human Infectiology MDx | Catalogue 2011 - Diagenode Diagnostics

Herpes simplex 1 /Herpes simplex 2Ref: Dia-HSV1/2-050Herpes simplex viruses 1 and 2 (HSV-1 and HSV-2) are membersof the herpes virus family, Herpesviridae, which cause infectionsin humans (Ryan and Ray, 2004). All viruses in the herpes familyproduce life-long infections.These viruses, also known as Human Herpes Virus 1 and 2 (HHV-1and HHV-2) are neurotropic and neuroinvasive viruses; they enterand latent in the human nervous system, accounting for theirdurability in the human body. HSV-1 is commonly associated withherpes outbreaks of the face known as cold sores or fever blisters,whereas HSV-2 is more often associated with genital herpes.Ryan KJ, Ray CG (editors) (2004). Sherris Medical Microbiology (4th ed. ed.).McGraw Hill. pp. 555–62.VALIDATIONResearched pathogens:• Herpes simplex virus 1: FAM dye, emission: 520 nm• Herpes simplex virus 2: yellow dye, emission: 549 nmTarget genes: • Glycoprotein B (UL 27, Herpes simplex virus 1)• Glycoprotein B (UL 27, Herpes simplex virus 2)Number of wells by test: 1 (Herpes simplex virus 1/ Herpes simplex virus 2)Samples used:• Bronchoalveolar lavages• Tissue/biopsy• Vesicular fluid• Ocular fluid• Swab corneal scraping• Cerebrospinal fluid• Amniotic fluid• Plasma• SerumValidated extraction methods: • Nuclisens easyMAG ® System (Biomerieux)• MagNa pure LC system (Roche)• QIAamp ® UltraSens Virus Kit (Qiagen)• QiaSymphony (Qiagen)Real-time PCR systems: • ABI (7000/7300/7500/7900)• Roche LightCycler ® (480/2.0 on request)• Biorad (iCycler/IQ5/CFX96)• Qiagen Rotor-Gene (6000)• Stratagene (MX3000P/3005P)• Cepheid (SmartCycler II)PCR volumes: 50 µl, 25 µl (depending of the real-time PCR system used)PCR reactions/kit:• 50 reactions (50 µl PCR, final volume)• 100 reactions (25 µl PCR, final volume)In combination with:• Universal inhibition control (DIA-UIC-050):DIA-UIC(DR)-050: orange dye, emission: 576 nmDIA-UIC(TR)-050: texas red dye, emission: 603 nmDIA-UIC(Cy5)-050: Cy5 dye, emission: 662 nm• DNA extraction & PCR inhibition control (DIA-EIC/DNA)-050:DIA-EIC/DNA(DR)-050: orange dye, emission: 576 nmDIA-EIC/DNA(TR)-050: texas red dye, emission: 603 nmDIA-EIC/DNA(Cy5)-050: Cy5 dye, emission: 662 nm23